Adipose Tissue-Derived Microvascular Fragments From Male and Female Fat Donors Exhibit a Comparable Vascularization Capacity

Adipose tissue-derived microvascular fragments (MVF) represent effective vascularization units for tissue engineering. Most experimental studies exclusively use epididymal fat tissue of male donor mice as a source for MVF isolation. However, in future clinical practice, MVF-based approaches may be applied in both male and female patients. Therefore, we herein compared the vascularization capacity of MVF isolated from the epididymal and peri-ovarian fat tissue of male and female donor mice. Freshly isolated MVF from male and female donors did not differ in their number, length distribution, viability and cellular composition. After their assembly into spheroids, they also exhibited a comparable in vitro sprouting activity. Moreover, they could be seeded onto collagen-glycosaminoglycan matrices, which were implanted into full-thickness skin defects within mouse dorsal skinfold chambers. Repetitive intravital fluorescence microscopy as well as histological and immunohistochemical analyses revealed a comparable vascularization and incorporation of implants seeded with MVF of male and female origin. Taken together, these findings demonstrate that the vascularization capacity of MVF is not gender-specific.

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Vascularization of Microvascular Fragment Isolates from Visceral and Subcutaneous Adipose Tissue of Mice

Tissue Engineering and Regenerative Medicine ◽

10.1007/s13770-021-00391-8 ◽

2021 ◽

Author(s):

Thomas Später ◽

Julia E. Marschall ◽

Lea K. Brücker ◽

Ruth M. Nickels ◽

Wolfgang Metzger ◽

...

Keyword(s):

Adipose Tissue ◽

Connective Tissue ◽

Visceral Fat ◽

Subcutaneous Fat ◽

Experimental Studies ◽

Length Distribution ◽

Epididymal Adipose Tissue ◽

Fat Tissue ◽

Subcutaneous Fat Tissue

Abstract Background: Adipose tissue-derived microvascular fragments (MVF) represent effective vascularization units for tissue engineering. Most experimental studies in rodents exclusively use epididymal adipose tissue as a visceral fat source for MVF isolation. However, in future clinical practice, MVF may be rather isolated from liposuctioned subcutaneous fat tissue of patients. Therefore, we herein compared the vascularization characteristics of MVF isolates from visceral and subcutaneous fat tissue of murine origin. Methods: MVF isolates were generated from visceral and subcutaneous fat tissue of donor mice using two different enzymatic procedures. For in vivo analyses, the MVF isolates were seeded onto collagen-glycosaminoglycan scaffolds and implanted into full-thickness skin defects within dorsal skinfold chambers of recipient mice. Results: By means of the two isolation procedures, we isolated a higher number of MVF from visceral fat tissue when compared to subcutaneous fat tissue, while their length distribution, viability and cellular composition were comparable in both groups. Intravital fluorescence microscopy as well as histological and immunohistochemical analyses revealed a significantly reduced vascularization of implanted scaffolds seeded with subcutaneous MVF isolates when compared to implants seeded with visceral MVF isolates. Light and scanning electron microscopy showed that this was due to high amounts of undigested connective tissue within the subcutaneous MVF isolates, which clogged the scaffold pores and prevented the interconnection of individual MVF into new microvascular networks. Conclusion: These findings indicate the need for improved protocols to generate connective tissue-free MVF isolates from subcutaneous fat tissue for future translational studies.

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Cellular adaptations in fat tissue of exercise-trained miniature swine: role of excess energy intake

Journal of Applied Physiology ◽

10.1152/jappl.2000.88.3.881 ◽

2000 ◽

Vol 88 (3) ◽

pp. 881-887 ◽

Cited By ~ 5

Author(s):

Gale B. Carey

Keyword(s):

Adipose Tissue ◽

Energy Intake ◽

Excess Energy ◽

Fat Tissue ◽

Cellular Mechanisms ◽

Miniature Swine ◽

Extracellular Adenosine

This study examined the influence of energy expenditure and energy intake on cellular mechanisms regulating adipose tissue metabolism. 1 Twenty-four swine were assigned to restricted-fed sedentary, restricted-fed exercise-trained, full-fed sedentary, or full-fed exercise-trained groups. After 3 mo of treatment, adipocytes were isolated and adipocyte size, adenosine A1 receptor characteristics, and lipolytic sensitivity were measured. Swine were infused with epinephrine during which adipose tissue extracellular adenosine, plasma fatty acids, and plasma glycerol were measured. Results revealed that adipocytes isolated from restricted-fed exercised swine had a smaller diameter, a lower number of A1 receptors, and a greater sensitivity to lipolytic stimulation, compared with adipocytes from full-fed exercised swine. Extracellular adenosine levels were transiently increased on infusion of epinephrine in adipose tissue of restricted-fed exercised but not full-fed exercised swine. These results suggest a role for adenosine in explaining the discrepancy between in vitro and in vivo lipolysis findings and underscore the notion that excess energy intake dampens the lipolytic sensitivity of adipocytes to β-agonists and adenosine, even if accompanied by exercise training.

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Sex difference in insulin-stimulated glucose transport in rat and human adipocytes

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1984.246.3.e211 ◽

1984 ◽

Vol 246 (3) ◽

pp. E211-E215 ◽

Cited By ~ 3

Author(s):

J. E. Foley ◽

A. Kashiwagi ◽

H. Chang ◽

T. P. Huecksteadt ◽

S. Lillioja ◽

...

Keyword(s):

Adipose Tissue ◽

Glucose Transport ◽

Subcutaneous Tissue ◽

Subcutaneous Fat ◽

Transport Rate ◽

Female Rats ◽

Fat Tissue ◽

Male And Female ◽

Male And Female Rats ◽

Female Subjects

In an effort to determine whether differences in basal and maximum insulin-stimulated glucose transport by isolated adipocytes are a function of donor sex, we measured glucose transport rates in the absence and presence of 8 nM insulin in adipocytes isolated from the abdominal subcutaneous fat tissue of nine male and ten female subjects with varying degrees of obesity and in adipocytes isolated from the abdominal subcutaneous and retroperitoneal fat tissue of (180-220 g) male and female rats. Because maximal insulin-stimulated glucose transport rate per cell of adipocytes isolated from subcutaneous abdominal tissue of male and female subjects was constant in each sex, the data have been normalized on the basis of transport per cell. The results demonstrated that basal and maximal insulin-stimulated glucose transport per cell was 53-75% higher per cell in the females versus males in adipocytes from human subcutaneous abdominal adipose tissue (P less than 0.01). A similar difference in glucose transport rate between males and females (P less than 0.001) was also found in rat abdominal subcutaneous adipose tissue. Adipocytes isolated from rat retroperitoneal adipose tissue had higher transport rates (approximately three-fold) and smaller sex differences (35% higher in females) than found in adipocytes from rat and human subcutaneous tissue. These results indicate that basal and maximum insulin-stimulated glucose transport is higher by adipocytes isolated from females and that this difference is independent of adipose cell size and species.

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OPTICAL COHERENCE TOMOGRAPHY OF ADIPOSE TISSUE AT PHOTODYNAMIC/PHOTOTHERMAL TREATMENT IN VITRO

Journal of Innovative Optical Health Sciences ◽

10.1142/s1793545813500107 ◽

2013 ◽

Vol 06 (02) ◽

pp. 1350010 ◽

Cited By ~ 3

Author(s):

IRINA YU. YANINA ◽

NATALIA A. TRUNINA ◽

VALERY V. TUCHIN

Keyword(s):

Adipose Tissue ◽

Brilliant Green ◽

Light Exposure ◽

Fat Tissue ◽

Fat Cell ◽

Refractive Index Distribution ◽

Tissue Slices ◽

Photothermal Treatment ◽

Index Distribution

Temporal changes in structure and refractive-index distribution of adipose tissue at photodynamic/photothermal treatment were studied with OCT in vitro. Ethanol–water solutions of indocyanine green (ICG) and brilliant green (BG) were used for fat tissue staining. CW laser diode (808 nm) and LED light source (442 and 597 nm) were used for irradiation of stained tissue slices. The data received supporting the hypothesis that photodynamic/photothermal treatment, induces fat cell lipolysis during a certain period of time after light exposure.

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Adipose tissue can be generated in vitro by using adipocytes from human fat tissue mesenchymal stem cells seeded and cultured on fibrin gel sheet

Cell and Tissue Banking ◽

10.1007/s10561-012-9304-6 ◽

2012 ◽

Vol 14 (1) ◽

pp. 97-106 ◽

Cited By ~ 2

Author(s):

Cong Toai Tran ◽

Duy Thao Huynh ◽

Ciro Gargiulo ◽

Le Bao Ha Tran ◽

Minh Hang Huynh ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Fat Tissue ◽

Fibrin Gel

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The Fabp4-Cre-Model is Insufficient to Study Hoxc9 Function in Adipose Tissue

Biomedicines ◽

10.3390/biomedicines8070184 ◽

2020 ◽

Vol 8 (7) ◽

pp. 184

Author(s):

Sebastian Dommel ◽

Claudia Berger ◽

Anne Kunath ◽

Matthias Kern ◽

Martin Gericke ◽

...

Keyword(s):

Body Weight ◽

Adipose Tissue ◽

High Fat Diet ◽

Fat Distribution ◽

Chow Diet ◽

Littermate Control ◽

Male And Female ◽

Biologically Relevant

Developmental genes are important regulators of fat distribution and adipose tissue (AT) function. In humans, the expression of homeobox c9 (HOXC9) is significantly higher in subcutaneous compared to omental AT and correlates with body fat mass. To gain more mechanistic insights into the role of Hoxc9 in AT, we generated Fabp4-Cre-mediated Hoxc9 knockout mice (ATHoxc9-/-). Male and female ATHoxc9-/- mice were studied together with littermate controls both under chow diet (CD) and high-fat diet (HFD) conditions. Under HFD, only male ATHoxc9-/- mice gained less body weight and exhibited improved glucose tolerance. In both male and female mice, body weight, as well as the parameters of glucose metabolism and AT function were not significantly different between ATHoxc9-/- and littermate control CD fed mice. We found that crossing Hoxc9 floxed mice with Fabp4-Cre mice did not produce a biologically relevant ablation of Hoxc9 in AT. However, we hypothesized that even subtle reductions of the generally low AT Hoxc9 expression may cause the leaner and metabolically healthier phenotype of male HFD-challenged ATHoxc9-/- mice. Different models of in vitro adipogenesis revealed that Hoxc9 expression precedes the expression of Fabp4, suggesting that ablation of Hoxc9 expression in AT needs to be achieved by targeting earlier stages of AT development.

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Secretome of Adipose Tissue-Derived Stem Cells (ASCs) as a Novel Trend in Chronic Non-Healing Wounds: An Overview of Experimental In Vitro and In Vivo Studies and Methodological Variables

International Journal of Molecular Sciences ◽

10.3390/ijms20153721 ◽

2019 ◽

Vol 20 (15) ◽

pp. 3721 ◽

Cited By ~ 8

Author(s):

Francesca Lombardi ◽

Paola Palumbo ◽

Francesca Rosaria Augello ◽

Maria Grazia Cifone ◽

Benedetta Cinque ◽

...

Keyword(s):

Wound Healing ◽

Adipose Tissue ◽

Experimental Studies ◽

Extrinsic Factors ◽

Experimental Conditions ◽

In Vivo Models ◽

Large Variability ◽

Healing Wounds

Wound healing is a complex process with a linear development that involves many actors in a multistep timeline commonly divided into four stages: Hemostasis, inflammation, proliferation, and remodeling. Chronic non-healing wounds fail to progress beyond the inflammatory phase, thus precluding the next steps and, ultimately, wound repair. Many intrinsic or extrinsic factors may contribute to such an occurrence, including patient health conditions, age-related diseases, metabolic deficiencies, advanced age, mechanical pressure, and infections. Great interest is being focused on the adipose tissue-derived stem cell’s (ASC) paracrine activity for its potential therapeutic impact on chronic non-healing wounds. In this review, we summarize the results of in vitro and in vivo experimental studies on the pro-wound healing effects of ASC-secretome and/or extracellular vesicles (EVs). To define an overall picture of the available literature data, experimental conditions and applied methodologies are described as well as the in vitro and in vivo models chosen in the reported studies. Even if a comparative analysis of the results obtained by the different groups is challenging due to the large variability of experimental conditions, the available findings are undoubtedly encouraging and fully support the use of cell-free therapies for the treatment of chronic non-healing wounds.

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The Differences in the Characteristics of Insulin-producing Cells Using Human Adipose-tissue Derived Mesenchymal Stem Cells from Subcutaneous and Visceral Tissues

Scientific Reports ◽

10.1038/s41598-019-49701-0 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Yuma Wada ◽

Tetsuya Ikemoto ◽

Yuji Morine ◽

Satoru Imura ◽

Yu Saito ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Insulin Secretion ◽

Fat Tissue ◽

Adipose Tissues ◽

Insulin Producing Cells ◽

Proliferation Ability ◽

Visceral Adipose

Abstract The aim of this study was to investigate the characteristics of insulin producing cells (IPCs) differentiated from adipose-tissue derived stem cells (ADSCs) isolated from human subcutaneous and visceral adipose tissues and identify ADSCs suitable for differentiation into efficient and functional IPCs. Subcutaneous and visceral adipose tissues collected from four (4) patients who underwent digestive surgeries at The Tokushima University (000035546) were included in this study. The insulin secretion of the generated IPCs was investigated using surface markers by: fluorescence activated cell sorting (FACS) analysis; cytokine release; proliferation ability of ADSCs; in vitro (glucose-stimulated insulin secretion: (GSIS) test/in vivo (transplantation into streptozotocin-induced diabetic nude mice). The less fat-related inflammatory cytokines secretions were observed (P < 0.05), and the proliferation ability was higher in the subcutaneous ADSCs (P < 0.05). Insulin expression and GISI were higher in the subcutaneous IPCs (P < 0.01 and P < 0.05, respectively). The hyperglycaemic state of all mice that received IPCs from subcutaneous fat tissue converted into normo-glycaemia in thirty (30) days post-transplantation (4/4,100%). Transplanted IPCs were stained using anti-insulin and anti-human leukocyte antigen antibodies. The IPCs generated from the ADSCs freshly isolated from the human fat tissue had sufficient insulin secreting ability in vitro and in vivo.

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Antiretroviral-Related Adipocyte Dysfunction and Lipodystrophy in HIV-Infected Patients: Alteration of the PPARγ-Dependent Pathways

PPAR Research ◽

10.1155/2009/507141 ◽

2009 ◽

Vol 2009 ◽

pp. 1-8 ◽

Cited By ~ 19

Author(s):

Martine Caron ◽

Corinne Vigouroux ◽

Jean-Philippe Bastard ◽

Jacqueline Capeau

Keyword(s):

Adipose Tissue ◽

Cell Function ◽

Nuclear Translocation ◽

Fat Tissue ◽

Related Function ◽

Adipose Cell ◽

Cell Dysfunction ◽

Mrna And Protein Expression ◽

A Cell

Lipodystrophy and metabolic alterations are major complications of antiretroviral therapy in HIV-infected patients. In vitro studies using cultured murine and human adipocytes revealed that some protease inhibitors (PIs) and nucleoside reverse transcriptase inhibitors (NRTIs) were implicated to a different extent in adipose cell dysfunction and that a chronic incubation with some PIs decreased mRNA and protein expression of PPARγ. Defective lamin A maturation linked to PI inhibitory activity could impede the nuclear translocation of SREBP1c, therefore, reducing PPARγexpression. Adipose cell function was partially restored by the PPARγagonists, thiazolidinediones. Adverse effects of PIs and NRTIs have also been reported in macrophages, a cell type that coexists with, and modulates, adipocyte function in fat tissue. In HIV-infected patients under ART, a decreased expression of PPARγand of PPARγ-related genes was observed in adipose tissue, these anomalies being more severe in patients with ART-induced lipoatrophy. Altered PPARγexpression was reversed in patients stopping PIs. Treatment of patients with agonists of PPARγcould improve, at least partially, the subcutaneous lipoatrophy. These data indicate that decreased PPARγexpression and PPARγ-related function, resulting from ART-induced adipose tissue toxicity, play a central role in HIV-related lipoatrophy and metabolic consequences.

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Experimental Studies on a Recombinant Hirudin, CGP 39393

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648151 ◽

1991 ◽

Vol 65 (04) ◽

pp. 355-359 ◽

Cited By ~ 12

Author(s):

E Gray ◽

J Watton ◽

S Cesmeli ◽

T W Barrowcliffe ◽

D P Thomas

Keyword(s):

Thrombin Generation ◽

Bleeding Time ◽

Thrombus Formation ◽

Experimental Studies ◽

Venous Stasis ◽

Antithrombotic Agent ◽

Recombinant Hirudin ◽

Excessive Bleeding ◽

Generation Test

SummaryThe in vitro anticoagulant activities of recombinant desulphatohirudin (r-hirudin) were studied in the activated partial thromboplastin time (APTT) and the thrombin generation test : systems. In the APTT at concentrations below 5 μg/ml, r-hirudin showed a dose-response curye. At concentrations above 5 μg/ml, the plasma became unclottable, but in the thrombin generation test , at least 10 μg/ml of r-hirudin was required for full inhibition of thrombin generation. The antithrombotic effect was assessed using a rabbit venous stasis model; 150 μg/ml r-hirudin completely prevented thrombus formation at 10 and 20 min stasis. At antithrombotic dose, the mean bleeding time ratio measured in a rabbit ear template model, was not prolonged over control values. At higher doses, the bleeding time ratios were higher than those observed for the same dosage of heparin. These data indicate that while r-hirudin is an effective antithrombotic agent, antithrombotic doses have to be carefully titrated to avoid excessive bleeding.

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